(A) Location of the studied ACTA1 mutations on the 3D-representation of the actin molecule . Mutations indicated in green were biochemically analysed in this study, mutations indicated in pink were analysed in Costa et al. . (B) Native gel analysis of twelve α-actin mutants causing NM. Autoradiograms of native gels of 35S-labeled WT α-actin and 12 mutants causing NM run with or without ATP. Actins were produced in in vitro transcription translation reactions in reticulocyte lysate, which endogenously contains CAP  and the actin folding machinery, prefoldin (PFD) and CCT [8, 9, 15]. The positions of these complexes with actin are indicated. The graph underneath indicates the percentage of actin in each of the bands (associated with CCT (purple), with prefoldin (white, (+ATP)) or CAP (white, (-ATP)) and free actin (red). In each lane the total actin is the sum of these three actin species.